巴西牵牛的组织培养技术研究

以在试管中萌发的巴西牵牛种子苗的茎尖为外殖体，对巴西牵牛的组织培养技术进行了研究。茎尖生长成完整的试管植株和腋芽生长增殖的最佳培养基为MS＋IBA0．2mg／1。巴西牵牛试管苗驯化移栽的最佳基质为上面1／4蛭石下面3／4腐质土的双层基质。     

红叶石楠的组织培养研究

取红叶石楠的不同器官、不同部位的材料进行离体培养及快速繁殖研究。试验结果表明,根部萌发蘖条和盆栽大苗的嫩茎为首选外植体,在MS+6-BA 2mg.L-1(单位下同)+NAA0.5的培养基上有较高的腋芽萌发率,增殖阶段以MS+6-BA 4+NAA0.1培养基上的小苗最多,生长健壮,生根在1/2MS+IBA 0.25的培养基上进行。并且分析了生根较低的原因及提出改进措施。     

淡黄花百合的离体保存

对淡黄花百合组培苗的离体保存研究结果表明:在培养温度15-24℃,光照时间11h/d,光照强度1500lx条件下,最适培养基为添加1-3%甘露醇的MS培养基,保存10个月后存活率为92%左右,存活的幼苗接种于增殖培养基上,100%正常生长和增殖。     

Microstructure,Content and in vitro Release of Brucine and Strychnine in Strychnos Nux-Vomica Powder with Different Particle Sizes

To explore the effect of particle size on the quality uniformity and in vitro release performance of Strychnosnux-vomica powder, seven samples of Strychnos nux-vomica powder with different particle sizes were prepared.Microstructures and particle sizes were analyzed, and high performance liquid chromatography （HPLC） was used totest the contents and in vitro release performances of brucine and strychnine in the samples. Results showed that thecontents and the in vitro release rates of brucine （or strychnine） in different samples were different since there aredifferent proportions of endosperms to epidermal cells in Strychnos nux-vomica powder with different particle sizes.Brucine and strychnine in each sample were promptly released in the first ten minutes and their cumulative releaserates were higher than 70% after ten minutes. Eighty minutes later, the cumulative release rate tended to be a constant.Considering the quality uniformity and safety of Strychnos nux-vomica powder used as traditional Chinese medicine, itwould be better to control the particle size of Strychnos nux-vomica powder between 100 and 140 mesh in which themaximum cumulative release rate in vitro of brucine and strychnine can be relatively low within this range.     

Comparative effects of plant growth regulators on leaf and stem explants of Labisia pumila var. alata

Objective: Labisia pumila var. alata, commonly known as ’Kacip Fatimah’ or ’Selusuh Fatimah’ in Southeast Asia, is traditionally used by members of the Malay community because of its post-partum medicinal properties. Its various pharmaceutical applications cause an excessive harvesting and lead to serious shortage in natural habitat. Thus, this in vitro propagation study investigated the effects of different plant growth regulators (PGRs) on in vitro leaf and stem explants of L. pumila. Methods: The capabilities of callus, shoot, and root formation were evaluated by culturing both explants on Murashige and Skoog (MS) medium supplemented with various PGRs at the concentrations of 0, 1, 3, 5, and 7 mg/L. Results: Medium supplemented with 3 mg/L indole-3-butyric acid (IBA) showed the optimal callogenesis from both leaf and stem explants with (72.34±19.55)% and (70.40±14.14)% efficacy, respectively. IBA was also found to be the most efficient PGR for root induction. A total of (50.00±7.07)% and (77.78±16.47)% of root formation were obtained from the in vitro stem and leaf explants after being cultured for (26.5±5.0) and (30.0±8.5) d in the medium supplemented with 1 and 3 mg/L of IBA, respectively. Shoot formation was only observed in stem explant, with the maximum percentage of formation ((100.00±0.00)%) that was obtained in 1 mg/L zeatin after (11.0±2.8) d of culture. Conclusions: Callus, roots, and shoots can be induced from in vitro leaf and stem explants of L. pumila through the manipulation of types and concentrations of PGRs.     

临床分离猪链球菌对常用抗菌药的敏感性

以金黄色葡萄球菌ATCC25923作为质控菌,用微量稀释法测定了7种药物对临床分离猪链球菌的体外最小抑菌浓度(MIC),以临床检验标准委员会(CLSI)的临界浓度作为判断标准,判定了猪链球菌对7种药物的耐药性;并分别在外排泵抑制剂氰氯苯腙(CCCP)、利血平存在的情况下,测定红霉素、泰乐菌素对分离菌MIC的影响.微量稀释法测定的MIC结果显示:临床分离的菌株以耐药菌为主,耐药谱型达11种;对磺胺间甲氧嘧啶(100%)耐药性最为严重,对红霉素(39.5%)、泰乐菌素(31.6%)、恩诺沙星(28.9%)、环丙沙星(28.9%)耐药性次之,而氟苯尼考和氨苄西林对所选菌株均敏感.在外排泵抑制剂存在时,抗菌药物对部分分离的耐药菌MIC下降,提示猪链球菌存在与抗菌药物耐药相关的主动外排机制.以上结果提示,国内分离的猪链球菌对临床常用的药物产生一定的耐药性,在临床选用药物进行治疗时,应充分考虑菌株的耐药性问题,以免延误病情造成治疗失败.     

海枫藤花粉活力的测定方法比较

花粉在植物有性生殖过程中有重要作用．解析花粉发育、花粉活力等过程是当前研究的热点问题．研究结果表明：I—KI染色法、醋酸洋红染色法、TFC染色法不适用于测定海枫藤花粉的生活力,离体萌发法是测定海枫藤花粉生活力的最佳方法．     

大鼠大脑皮层星形胶质细胞体外培养方法建立及鉴定

目的:纯化和鉴定体外培养的大鼠大脑皮层星形胶质细胞.方法:取1~3天新生大鼠大脑皮层细胞进行体外培养,通过差速贴壁和传代去除成纤维细胞和神经元等,采用免疫组化法检测星形胶质细胞特异性蛋白—胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)鉴定星形胶质细胞.结果:培养的细胞具有典型的星形胶质细胞形态且生长旺盛,第3代95%以上为星形胶质细胞,特异性抗原GFAP表达阳性.结论:该方法操作简单,可靠,是一种有效的新生大鼠大脑皮层星形胶质细胞体外培养方法.     

Effects of the crude extract of Polygala tenuifolia Willd on human sperm in vitro

The aim of the present study is to analyze sperm membrane changes and the spermicidal effect in treatment with the crude extract from Polygala tenuifolia Willd (PTW) in vitro. The root of PTW was extracted in distilled water. Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer assay) of the extract from the PTW root. The hypo-osmotic swelling (HOS) test and the eosin Y (EY) staining were used to detect the integrity of sperm membrane and vitality. The sperm chromatin dispersion (SCD) test was performed to determine sperm DNA integrity. N-9 was used as a reference standard and semen added to physiological saline was used as the control. Semen samples were donated by 42 healthy fertile men. The crude extract from the root of PTW could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 20.0 and 10.0 mg/ml; at the concentration of 5.0 mg/ml, spermatozoa were immobilized in (39.5±3.2) s. In the groups of the crude extract from the root of PTW and N-9 solution, the rate of the normal HOS (tails swollen) and the white head (unstained) was 0%, and the rate of the abnormal HOS (tails unswollen) and red head (stained) was 100%. Sperm DNA fragmentation showed no change in exposure to the crude extract from the root of PTW and N-9 solution. The sperm revival test did not show any spermatozoa that recovered their motilities. The rapid spermicidal activity of the crude extract from the root of PTW in vitro may occur by the disruption of the sperm membrane integrity.     

Normal epigenetic inheritance in mice conceived by in vitro fertilization and embryo transfer

An association between assisted reproductive technology (ART) and neurobehavioral imprinting disorders has been reported in many studies, and it seems that ART may interfere with imprint reprogramming. However, it has never been explored whether epigenetic errors or imprinting disease susceptibility induced by ART can be inherited transgenerationally. Hence, the aim of this study was to determine the effect of in vitro fertilization and embryo transfer (IVF-ET) on transgenerational inheritance in an inbred mouse model. Mice derived from IVF-ET were outcrossed to wild-type C57BL/6J to obtain their female and male line F2 and F3 generations. Their behavior, morphology, histology, and DNA methylation status at several important differentially methylated regions (DMRs) were analyzed by Morris water maze, hematoxylin and eosin (H&E) staining, and bisulfite genomic sequencing. No significant differences in spatial learning or phenotypic abnormality were found in adults derived from IVF (F1) and female and male line F2 and F3 generations. A borderline trend of hypomethylation was found in H19 DMR CpG island 3 in the female line-derived F3 generation (0.40±0.118, P=0.086). Methylation status in H19/Igf2 DMR island 1, Igf2 DMR, KvDMR, and Snrpn DMR displayed normal patterns. Methylation percentage did not differ significantly from that of adults conceived naturally, and the expression of the genes they regulated was not disturbed. Transgenerational integrity, such as behavior, morphology, histology, and DNA methylation status, was maintained in these generations, which indicates that exposure of female germ cells to hormonal stimulation and gamete manipulation might not affect the individuals and their descendents.